Source: Los Gatos Research submitted to NRP
LASER-BASED SENSOR FOR NITRATE ISOTOPE ANALYSIS IN WATER POLLUTION MONITORING
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
COMPLETE
Funding Source
Reporting Frequency
Annual
Accession No.
1000295
Grant No.
2013-33610-21304
Cumulative Award Amt.
$450,000.00
Proposal No.
2013-03544
Multistate No.
(N/A)
Project Start Date
Sep 1, 2013
Project End Date
Nov 30, 2017
Grant Year
2013
Program Code
[8.4]- Air, Water and Soils
Recipient Organization
Los Gatos Research
67 East Evelyn Avenue, Suite 3
Mountain View,CA 94041
Performing Department
(N/A)
Non Technical Summary
In this Small Business Innovative Research (SBIR) effort, Los Gatos Research (LGR) proposes to utilize its patented Off-Axis ICOS technology to develop an analyzer for the accurate determination of 15N/14N, 18O/16O, and 17O/16O isotope ratios in nitrates (d15N, d18O, and d17O). The system will employ the bacterial denitrification method to convert nitrates (NO3-) into N2O, and then utilize a mid-infrared Off-Axis ICOS analyzer to rapidly (< 5 minutes) determine nitrate concentration, d15N, d18O, and d17O to better than ± 1%, ± 0.2 ‰, ± 0.5 ‰, and ± 0.5 ‰ respectively. Nitrate contamination in water is an ubiquitous, world-wide problem, and the resulting instrument will help provide nitrate source apportionment, identify multiple nitrate sources, study spatial mixing of nitrate pollution, and identify areas in which natural nitrate attenuation processes are taking place. Additionally, the SBIR analyzer can be used to help quantify the nitrate pollution and N2O emission impacts of emerging biofuels. In Phase I, LGR demonstrated technical feasibility by fabricating a prototype system for quantification of N2O, 15N14NO, 14N15NO, and 14N14N18O in digested nitrate samples. The prototype was tested on a variety of gas cylinders and found to quantify [N2O], d15N, d15Na, d15Nb, and d18O with a precision of ±0.01 ppb, ±0.42 ‰, ±0.40 ‰, ±0.50 ‰, and ±0.75 ‰ respectively (1s, 1000s for [N2O] = 320 ppb). Repeated measurements of discrete reference gas injections demonstrated that the Phase I sensor was precise to better than ±0.25 ‰ and ±0.53 ‰ for d15N and d18O respectively with minimal dependence of the isotope ratio on nitrous oxide concentration over a wide dynamic range of [N2O] = 0.25 - 2 ppm. Potential cross-interferences with ambient water vapor and higher hydrocarbons were mitigated by using front-end cold trap. The instrument was then used to characterize nitrate-polluted water samples obtained from the Abbotsford aquifer (British Columbia, Canada). The Phase I system was directly compared to isotope ratio mass spectrometry (IRMS) measurements and found to be accurate to within ±2.4 ‰ and ±3.2 ‰ for d15N and d18O. This data clearly showed that the high levels of nitrate pollution were due to animal waste and fertilizer ammonium runoff and not to atmospheric deposition or fertilizer nitrate runoff. Due to the novelty of the Phase I measurements, they will be presented at the European Geophysical Union Spring Meeting (Vienna, Austria) and the American Geophysical Union Fall Meeting (San Francisco, California). Finally, the results were also used to help identify Phase II improvements. In Phase II, Los Gatos Research will refine the Nitrate Isotope Analyzer hardware, electronics, and software to improve the instrument accuracy, precision, and stability. Then, the technology will be extended to include measurements of d17O. The instrument will be automated by including provisions for autonomous sample conversion and handing. Subsequent to extensive laboratory testing at LGR, the analyzer will be deployed at the International Atomic Energy Agency (Vienna, Austria) and tested by the Isotope Hydrology Laboratory at no charge to this SBIR effort. A second Nitrate Isotope Analyzer will be fabricated and deployed at Purdue University, where Professor Greg Michalski in the Department of Chemistry will verify its accuracy against an isotope ratio mass spectrometer (IRMS) and use it to quantify water pollution sources including atmospheric deposition, nitrate fertilizers, and soil runoff. During Phase III, LGR will sell the N2O isotope analyzers to isotope measurement laboratories, environmental research groups, global monitoring stations, and water quality management agencies. A preliminary market analysis suggests a 5-year commercial revenue exceeding $40M for these four markets alone. A complete Commercialization Plan is attached, including letters of support from potential customers and international distributors.
Animal Health Component
100%
Research Effort Categories
Basic
(N/A)
Applied
100%
Developmental
(N/A)
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
11202102000100%
Knowledge Area
112 - Watershed Protection and Management;

Subject Of Investigation
0210 - Water resources;

Field Of Science
2000 - Chemistry;
Goals / Objectives
In this Small Business Innovative Research (SBIR) effort, Los Gatos Research (LGR) proposes to utilize its patented Off-Axis ICOS technology to develop an analyzer for the accurate determination of 15N/14N, 18O/16O, and 17O/16O isotope ratios in nitrates (d15N, d18O, and d17O). The system will employ the bacterial denitrification method to convert nitrates (NO3-) into N2O, and then utilize a mid-infrared Off-Axis ICOS analyzer to rapidly (< 5 minutes) determine nitrate concentration, d15N, d18O, and d17O to better than ± 1%, ± 0.2 ‰, ± 0.5 ‰, and ± 0.5 ‰ respectively. Nitrate contamination in water is an ubiquitous, world-wide problem, and the resulting instrument will help provide nitrate source apportionment, identify multiple nitrate sources, study spatial mixing of nitrate pollution, and identify areas in which natural nitrate attenuation processes are taking place. Additionally, the SBIR analyzer can be used to help quantify the nitrate pollution and N2O emission impacts of emerging biofuels.
Project Methods
Los Gatos Research uses mid-infrared Off-Axis ICOS to quantify isotope ratios of nitrous oxide (N2O). Nitrate samples are housed in 20 mL vials and denitrifying bacteria (Pseudomonas aureofaciens) are added to quantitatively convert the nitrate into N2O. The vial headspace, which contains ~ 10 - 20 ppmv N2O/He, is directly injected into the instrument without any sample conditioning. Mid-Infrared cavity-enhanced tunable diode laser absorption spectroscopy (e.g. Off-Axis ICOS) is used to accurately measure the concentrations of N2O, 15N14NO, 14N15NO, and N218O. The relevant atomic isotope ratios (e.g. 15N/14N and 18O/16O) are then calculated from the measured molecular densities and the nitrate concentration in the water sample is determined from the N2O mixing ratio.

Progress 09/01/13 to 11/30/17

Outputs
Target Audience: Nothing Reported Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?The results have been presented in various conferences or seminars (including American Geophysical Union Fall Meetings, European Geophysical Union Spring Meetings, and Advances in Stable Isotope Techniques and Applications Workship). How have the results been disseminated to communities of interest? Nothing Reported What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? In Phase II, Los Gatos Research has developed an analyzer for accurate measurements of N2O, 15N14NO, 14N15NO, 14N14N18O, and 14N14N17O in digested nitrate samples. The analyzer was tested on a variety of gas cylinders and found to quantify [N2O], d15N, d15Na, d15Nb, d18O, and d17O with a precision of ±0.03 ppb, ±0.28 ‰, ±0.49 ‰, ±0.64 ‰, and ±3.1 ‰ respectively (1s, 100s for [N2O] = 1 ppm). Repeated measurements of discrete reference gas injections demonstrated that the analyzer was precise to better than ±0.26 ‰, ±0.33 ‰, ±0.22 ‰, and ±0.27 ‰ for d15Na, d15Nb, d18O, and d17O respectively (300s per measurement for [N2O] = 10 ppm). One of the analyzers has been tested with the International Atomic Energy Agency. It has been used to evaluate chemical method (Cd-azide method) for conversion of aqueous NO3 or NO2 to N2O. The measurements are in excellent agreement with expected values, and with comparative results obtained by IRMS. The other analyzer has been delivered to Professor Greg Michalski at Purdue University for verification. Automated sample handling has been implemented by integrating a gas autosampler with an N2O isotope analyzer. The analyzer is now capable of making stand-alone, autonomous measurements of multiple digested water samples. All the above achievements have been presented at American Geophysical Union Fall Meeting, European Geophysical Union Spring Meeting, and ASITA Conference.

Publications


    Progress 09/01/13 to 08/31/14

    Outputs
    Target Audience:Scientific community that studies the sourcing of nitrates Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? Nothing Reported How have the results been disseminated to communities of interest?Preliminary results presented at AGU Fall Meetings What do you plan to do during the next reporting period to accomplish the goals?The major remaining tasks are expected to be completed on the following schedule: Completion of measurements by Purdue University: March 2016 Analysis and publication of Purdue field measurements: June 2016 Integration with autosampler: May 2016 Evaluate sample handling/automated conversion methods: July 2016 Final report: August 2016

    Impacts
    What was accomplished under these goals? LGR has made substantial progress towards the completion of this Phase II SBIR, completing many of the tasks outlined in the contract. Specific accomplishments include: Fabricated Off-Axis ICOS Nitrate Isotope Analyzer LGR has fabricated two improved performance N2O isotopic analyzers for lab testing within LGR and field evaluation by subcontractors to this work. Notable improvements from this work include the use of higher numerical aperture collection optics to increase the intensity of light collected onto the detector (and SNR) by approximately a factor of 2. Line fitting parameters (esp. pressure width and position) have been improved for fits over a larger gas concentration range resulting in improved performance over a dynamic range of 320 ppb to 100 ppm of N2O. Extended the Instrument to Quantify d17O LGR has extended the scanning range of the laser to cover a nearby 14N14N17O absorption feature. Although the feature is significantly smaller than the other isotopomers, LGR achieved a precision of better than 1‰ in 300 second for a concentration of 10 ppm N2O. Alternative spectral regions were evaluated as part of this work and found to yield improved performance for some subsets of isotopes, however, the original region provided the best results when each of the 5 isotopic ratios (d15N, d15Na, d15Nb, d18O, d17O) are of interest. Evaluated the Performance of the Instrument in LGR's Laboratory for: Accuracy: Better than <0.2‰ for d15N, d15Na, d15Nb, d18O and < 0.5‰ for d17O in 300 sec, 10 ppm N2O when referenced hourly. Dynamic Range: 320ppb - 100ppm Linearity: R2 of >0.999 over 320ppb - 100ppm Precision: <0.2‰ for d15N, d15Na, d15Nb, d18O and < 0.5‰ for d17O in 300 sec, 10 ppm N2O Long Term Stability: <2‰ peak-to-peak drift in d15Na, d15Nb and <3‰ peak-to-peak drift in d17O in 24 hrs for a sample concentration of 1 ppm N2O. In the first quarter of 2015, LGR delivered an N2O isotopic analyzer to Len Wassenaar at the International Atomic Energy Agency (IAEA) for evaluation against IRMS. In July, 2015 LGR delivered an N2O isotopic analyzer with d17O capability to Greg Michalski (Purdue University) to quantify nitrate processing in agricultural fields

    Publications